Immunogenicity and Protective Efficacy against Murine Tuberculosis of a Prime-Boost Regimen with BCG and a DNA Vaccine Expressing ESAT-6 and Ag85A Fusion Protein
Figure 2
Expression, purification, and identification of recombinant fusion protein. E. coli BL21 (DE3) harboring pPro685A was cultured with IPTG. The expression (a) and the purification procession of r685A fusion protein (b) were confirmed by SDS-PAGE and Western blotting (c). Lane M: protein molecular size marker (kDa); lane a1: E. coli strain without IPTG; lane a2: E. coli strain 2 h after IPTG induction; lane a3: E. coli strain 4 h after IPTG induction; lane a4: E. coli strain 6 h after IPTG induction; lane b1: cell lysis; lane b2: fraction from Ni-NTA column after wash with denaturing binding buffer; lanes b3 and b4: fraction after wash with wash buffer; lane b5: r685A protein eluted with native elution buffer; lane c1: anti-ESAT-6 antibody; lane c2: anti-Ag85A antibody.
